|Publication number||US7676071 B2|
|Application number||US 11/367,522|
|Publication date||Mar 9, 2010|
|Filing date||Mar 3, 2006|
|Priority date||Mar 4, 2005|
|Also published as||CN1869649A, CN1869649B, DE102006001427A1, DE502006002858D1, EP1698928A1, EP1698928B1, US20060232843|
|Publication number||11367522, 367522, US 7676071 B2, US 7676071B2, US-B2-7676071, US7676071 B2, US7676071B2|
|Original Assignee||Leica Microsystems Cms Gmbh|
|Export Citation||BiBTeX, EndNote, RefMan|
|Patent Citations (5), Classifications (6), Legal Events (2)|
|External Links: USPTO, USPTO Assignment, Espacenet|
Priority is claimed to German patent application DE 10 2005 010 490.8, filed Mar. 4, 2005, German patent application DE 10 2005 048 481.6 filed Oct. 7, 2005, and German patent application DE 10 2006 001 427.8 filed Jan. 10, 2006.
The present invention relates to a method for illuminating a specimen or a region of the specimen (region of interest, ROI), the information indicating whether and in what manner a specific point on the specimen is to be illuminated, being stored in a data memory under a specific address.
Methods for illuminating a specimen are used in the field, in particular in confocal microscopy. In this context, a specimen is scanned by a mostly laser-produced light beam in a line-by-line or meander pattern. However, in order to avoid damaging the specimen, only specific specimen regions, the regions of interest (ROIs), are illuminated. Since the light beam, conditionally upon the inertia of the light beam deflection, sweeps over the entire specimen, its intensity is modulated as a function of the position on the specimen. The information indicating whether a point of a specimen is to be illuminated is stored in a memory and is read out from the same on the basis of the light beam position. The control data are then fed to the light modulator.
If the illuminated specimen is made up of cells in an aqueous solution, for example, then the position, orientation, and shape of the specimen or of the ROI do not remain constant. The changes in the specimen are mapped by adapting the intensity data in the memory. In the process, however, it may be necessary to update large areas of the memory. However, since the process of updating the memory contents and reading out from the same must take place in parallel and in real time, rigorous demands are placed on the speed of the memory used.
This becomes especially problematic when specimens are to be scanned at a high frame rate. In such a case, the available memories quickly reach their performance limits and become very costly. In addition, very high-performance processors are needed for computing the new memory contents.
In a schematic representation,
An aspect of the present invention is to devise a method for illuminating a specimen or a region of a specimen, which will allow for spatial changes in the specimen or in parts of the specimen, while simultaneously placing as few demands as possible on the hardware used.
In an embodiment, the present invention provides a method for illuminating a specimen or a region of interest (ROI) of the specimen with a light source. The method includes storing information indicating whether and in what manner each of a plurality of image points on the specimen is to be illuminated in a data memory under a specific address. A change in at least one of the position, orientation and shape of the specimen resulting from movements inside the specimen is recorded as at least one image. A transformation of coordinates of the image points is derived from the at least one recorded image using an image processing system. Unique memory addresses are assigned to the transformed coordinates. The data memory is accessed on the basis of the memory addresses and the contents of the data memory are read out so as to control the light source.
Exemplary embodiments of the present invention are described in the following and schematically shown in the drawings, in which:
With the present invention there is no need to update the entire memory contents in order to update the parameters for controlling the light source. Rather, in accordance with the present invention, the changes in the specimen or in the ROI may be mapped by changing the transformation rules. In accordance with an embodiment of the present invention, these transformation rules are used to adapt the coordinates of the image points to be illuminated to the changes in the specimen or the ROI. Memory addresses, which may be used for accessing the data memory, may then be uniquely assigned to the transformed coordinates of the image points to be illuminated. Finally, the information for controlling the light source may be read out from the selected memory locations.
The change in the position, orientation and/or shape of the specimen advantageously results in a translation, a rotation, a compression and/or an expansion of the specimen itself. The shape of the specimen may change on its own. In such a case, the change in the ROI would be a reaction to the same.
The changes may preferably be reproduced by transforming the coordinates of the image points to be illuminated through the use of conformal mappings. In this context, translations, rotations and scalings are preferably used. By performing these transformations, the currently active specimen structure is generally transformed point for point into an image region of the ROI or into a reference image. In both cases, it is a question of reference regions or reference points that are mostly defined at the beginning of the measurement.
The method according to the present invention may advantageously be applied to specimens having a plurality of ROIs. The change in each individual ROI is reproduced by using a separate transformation rule, each individual transformation rule generally being differently parameterized and/or being able to be differently constructed. Thus, one transformation rule may merely include a translation, while another transformation rule is composed of a translation, a rotation, and a scaling.
To achieve a simple and practical mode of operation, the parameters of the individual transformations may preferably be adapted automatically, continuously and/or in real time to the changes in the specimen or in the ROI. Images of the specimen or of parts of the specimen could be captured in time sequence. The changes may be recorded using a camera or a scanner, for example. The recorded images are subsequently analyzed using an image processing system. The data are fed to a further system for determining the altered parameters of the transformation. The parameters are preferably stored in registers, whose contents may be used by the systems for transforming the coordinates of the points to be illuminated.
An embodiment of the invention provides for the data memory to be preferably subdivided into a plurality of memory areas, all or at least some of the ROIs of a specimen being mapped into separate memory areas. Preferably, the memory areas are not only logically, but also physically separated from one another. Thus, for each ROI, the individual transformations may be performed completely independently and, in particular, not as mutually blocking operations.
With regard to controlling the light source, on the one hand, the intensity may be changed; on the other hand, the spectrum and/or the shape of a light bundle emitted by the light source may be influenced. The individual parameters used for controlling the light source are stored in the data memory and are preferably not changed during the execution time or, at least, not for the duration of a few scanning cycles.
A change in the specimen or the ROI may result in overlapping ROIs. In such a case, a separate set of parameters for controlling the light source is defined for each ROI, and the value used for controlling the light source is ascertained by averaging or summing the individual values. In the summation operation, in particular, to avoid an unacceptably high light intensity, the individual parameters are limited to a range between a minimum and a maximum value.
To facilitate integration of a system for carrying out the method according to the present invention, it may be readily implemented in a CPLD or FPGA module having a plurality of internal memories. This makes it possible for the architecture of a high-performance unit to include a plurality of parallel processors. The process of transforming the coordinates of the points to be illuminated may be implemented very advantageously using digital signal processors which are likewise able to be integrated into the modules mentioned.
An embodiment of a method according to the present invention is schematically illustrated in an implementation in
With regard to other advantageous embodiments of the method according to the present invention, to avoid repetitive explanations, reference is made to the general portion of the specification and to the appended claims. Finally, it is particularly noted that the exemplary embodiment described above is merely intended for purposes of exemplifying the claimed teaching, but not for limiting it to such exemplary embodiment.
1. deflecting electronics of the laser beam
2. x-position of the laser beam
3. x-position of the laser beam
4. address generator
7. intensity data
12. affine mappings
14. x-position following translation
15. y-position following translation
17. x-position following translation and rotation
18. y-position following translation and rotation
20. x-position following the affine mappings
21. y-position following the affine mappings
|Cited Patent||Filing date||Publication date||Applicant||Title|
|US4907158||May 25, 1988||Mar 6, 1990||Carl-Zeiss-Stiftung||Method for performing work on cells of a cell culture and apparatus therefor|
|US6898004 *||Sep 23, 2002||May 24, 2005||Olympus Optical Co., Ltd.||Microscope system|
|US20030179445||Nov 5, 2001||Sep 25, 2003||Garrick Maenle||Cytological imaging systems and methods|
|EP1085292A1||Mar 30, 2000||Mar 21, 2001||Sapporo Breweries Ltd.||Coordinate transforming method in position setting means of observation device and observation device equipped with coordinate transforming means|
|WO2005017597A1||Aug 4, 2004||Feb 24, 2005||Leica Microsystems||Method and system for device-independent determination of coordinates of a point displayed by means of a microscope|
|Cooperative Classification||G02B21/008, G02B21/365|
|European Classification||G02B21/00M4A9, G02B21/36V|
|Jun 16, 2006||AS||Assignment|
Owner name: LEICA MICROSYSTEMS CMS GMBH,GERMANY
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:WIDZGOWSKI, BERND;REEL/FRAME:017799/0511
Effective date: 20060406
|Mar 14, 2013||FPAY||Fee payment|
Year of fee payment: 4